Parasitol Res (2008) 102:647–655
DOI 10.1007/s00436-007-0806-2
ORIGINAL PAPER
The life cycle of Stephanoprora aylacostoma n.sp.
(Digenea: Echinostomatidae), parasite of the threatened snail
Aylacostoma chloroticum (Prosobranchia, Thiaridae),
in Argentina
M. Ostrowski de Núñez & M. G. Quintana
Received: 31 October 2007 / Accepted: 9 November 2007 / Published online: 12 December 2007
# Springer-Verlag 2007
Abstract A new species, Stephanoprora aylacostoma is
described and its life cycle was resolved experimentally.
The prosobranch snail Aylacostoma chloroticum Hylton
Scott, collected in the Yacyretá dam, Province of Misiones,
Argentina, was found naturally infected with large-tailed
cercariae possessing a prepharyngeal body and corpuscles
in the excretory system but lacking collar spines. Meta-
cercariae, which encysted on the gills of experimentally
infected fishes Cnesterodon decemmaculatus (Jenyns) and
Poecilia reticulata (Peters) (Poecilidae), developed collar
spines after 10 days. Tetragonopterid fishes Moenckhausia
dichroura (Kner), Astyanax erythropterus (Holmberg) and
Hyphesobrycon serpae (Durbin in Eigenmann) were found
infected naturally. Sexually mature adults were recovered
from domestic chicks at day 7 post-exposure. Eggs shed in
chick faeces developed to miracidia within 13–15 days;
sporocysts were found on the gills of snails. The new
species differs from other species of the genus in its larger
eggs, in the smaller, slender body and smaller collar spines
of the adult and in the morphological and biological
features of the larval stages.
M. Ostrowski de Núñez (*)
Departamento de Ciencias Biológicas, Facultad de Ciencias
Exactas y Naturales, Universidad de Buenos Aires,
Ciudad Universitaria,
Pabellón II. 1428,
Buenos Aires, Argentina
e-mail: ostrowski@bg.fcen.uba.ar
M. G. Quintana
Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”,
Av. Angel Gallardo 470, (C1405DJR),
Buenos Aires, Argentina
Introduction
The occurrence of molluscs of the genus Aylacostoma Spix,
1827 (Prosobranchia, Thiaridae) in Argentina and Paraguay
was first reported by Hylton Scott (1951, 1954). In
Argentina, she described species living in freshwater
habitats near the rapids of Apipé, on the Paraná River,
between the cities of Ituzaingó (27°37′ S, 56°40′ W) and
Posadas (27°20′ S, 55°55′ W; Fig. 1). Half a century later,
these endemic species are threatened of extinction due to
the construction of the Yacyretá Hydroelectric power plant
(2,700,000 KW; flooded area, 1,720 km2). Of the five
species identified until 1993 (before impoundment), only
Aylacostoma chloroticum (Fig. 1) persists in two relictual
populations at the upstream section of the reservoir
(Quintana and Mercado-Laczkó 1997). A raise in the water
level of the dam that will be undertaken in the near future
may increase the extinction risk of the local snail
population. This fact led to the development of a
conservation program to propagate A. chloroticum snails
in captivity and reintroduce them into presumed adequate
locations downstream Yacyretá dam.
The examination of some A. chloroticum specimens
from the relictual populations revealed that they were
infected with an unknown fauna of larval trematodes. The
aim of this paper was to describe the life cycle of a new
species of the genus Stephanoprora Odhner, 1902, the most
frequent larval trematode in the snail population.
Materials and methods
Specimens of A. chloroticum of up to 37 mm in length were
collected from the Paraná River at Heller Peninsula, (27°20′
S, 55°55′ W) near Posadas City, Province of Misiones,
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Fig. 1 Map of South America showing the sampling area: the Paraná River and the lake created by the dam at the hydroelectric power station of
Yacyretá, including Aylacostoma chloroticum, scale 10 mm
Argentina. The snails were kept individually in vials with
20 ml of tap water and observed for the emergence of
cercariae. Immediately after shedding, some snails were
dissected to check for intramolluscan stages, whereas
emerging cercariae were exposed to laboratory-reared
Poecilia reticulata (Peters) and Cnesterodon decemmacu-
latus (Jenyns) and specimens of C. decemmaculatus
collected in the field. Some individuals of the latter were
used as control to rule out previous infections; their
dissection indicated that although they showed metacercar-
iae of Diplostomidae, the gills were free of parasites.
Experimentally infected fishes from natural and experimen-
tal sources were placed separately in small aquaria and fed
with commercial fish food.
Eight newly hatched unfed chicks (Gallus gallus
domesticus L) were force-fed mature metacercariae (n=
11–50) from experimentally infected fishes. The chicks
were housed in cages at room temperature and fed with oats
and water ad libitum. They were necropsied from 3 to
7 days post-exposure (pe) to recover the adults.
To determine the natural second intermediate host, fishes
belonging to the Tetragonopteridae, Moenckhausia
dichroura (Kner), Astyanax erythropterus (Holmberg) and
Hyphesobrycon serpae (Durbin in Eigenmann) were sam-
pled at different sites along the dam.
Eggs laid spontaneously by experimentally obtained
adults and collected from chick faeces were put on
depression slides and covered with coverslips to follow
daily development. At the same time, larger groups of eggs
were maintained in Petri dishes. Three to seven newly
Parasitol Res (2008) 102:647–655
emerged miracidia were exposed to each of eight laborato-
ry-reared A. chloroticum of around 10 mm in length, which
were placed in 10 ml of tap water. These snails were
maintained separately in small vials for 48 h and then held
together in an aerated aquarium, fed with commercial fish
food at room temperature (ranging between 13 and 25°C
during the study period) and dissected at 28, 43, 71, 136
and 240 days pe. Three snails were fixed for histological
sections, embedded in paraffine wax, cut at 7 μm, stained
with hematoxiline and eosine and mounted in Canada
balsam. Heleobia parchappei (d’Orbigny) collected at
Lujan River (Buenos Aires Province) and measuring up to
3 mm in size were exposed individually to two miracidia in
5 ml of tap water.
The life spans of miracidia and cercariae were deter-
mined by observing specimens placed in vials with 2 and
5 ml of tap water, respectively, three to five times per day.
Larval stages and adults were studied alive and as
permanent or temporary whole mounts. Adults were fixed
in nearly boiling 4% formalin; stained with alcoholic
hydrochloric carmine, dehydrated in an ethanol series,
cleared in creosote and mounted in Canada balsam. Some
adults were fixed in 96% ethanol for further molecular
analysis. All larval stages were fixed in nearly boiling
4% formalin; miracidia were mounted in Fauré liquid
(Langeron 1942); rediae and cercariae were stained as
described for adults or cleared in lactophenol and mounted
in glycerine jelly or mounted in formol without applying
pressure for measurements. All measurements (length ×
width) are given in micrometres, with the range followed
Parasitol Res (2008) 102:647–655
Fig. 2 Stephanoprora aylacostoma n.sp. a adult scale 200 μm; b collar, scale 50 μm; c mother redia at 42 days pe, scale 200 μm; d natural
daughter redia, scale 200 μm; e mother redia (in vivo) at 130 days pe, scale 100 μm; f body of cercaria, scale 100 μm; g cercaria, scale 500 μm
by the mean in parentheses; in the case of spines, the range
is followed by the mean, standard deviation and number of
spines measured in parentheses.
Figures were made with the aid of a camera lucida, and
details were added free-hand. Specimens were deposited in
the Parasitological Collection of the Museo Argentino de
Ciencias Naturales “Bernardino Rivadavia”, Buenos Aires,
Argentina (MACN-Pa).
In addition, the following material was studied for
comparison: (1) S. uruguayense vouchers No. 58/1-15,
175/1-12, 177/1-15, 181/1-16 from G. domesticus and 34/1-
7 from Larus dominicanus (Lichtenstein, 1823) deposited
in the Universidad Nacional del Comahue Bariloche,
Argentina, and additional specimens from personal collec-
tion (MON); (2) S. podicipei Etchegoin et Martorelli, 1997:
paratype No 3903 from Podiceps major (Boddaert, 1783)
and No. 4584 from Sterna hirundinacea Lesson, 1831
deposited in the Colección Helmintológica del Museo de
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Ciencias Naturales, La Plata, Argentina (CHMLP), and
specimens from personal collection of F. Cremonte; (3) S.
argentinensis (Sutton et al. 1982): paratypes No. 656 C and
644 C deposited in the CHMLP; (4) S. paradenticulata
(Nasir and Rodríguez 1969) from Himantopus himantopus
(Linnaeus, 1758), microslide No. 71152, and S. dentic-
ulata from G. domesticus of Nasir and Scorza (1968)
microslide No.63112, both deposited in the USA National
Parasite Collection, Beltsville, Maryland, USA (USNPC);
(5) S. denticulata from G. domesticus (n=1) and Larus
ridibundus L. (n=3) from personal collection of M. Køie
(Denmark).
Results
Stephanoprora aylacostoma n.sp. (Figs. 2a–g; Figs. 3a–k)
Adult (Figs. 2a–b; 3i–k)
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Fig. 3 a Freshly laid egg; b egg at 13 days pe; c miracidium, a–c,
same scale 50 μm; d sporocyst, scale 200 μm; e gill of snail showing
two sporocysts, scale 200 μm; f mother redia at 71 days pe, scale
500 μm; g cercaria in resting position, scale 250 μm; h metacercaria
Description
Measurements: based on 15 permanent mounted, egg-bearing
specimens recovered at day 7 pe from chicks exposed to cysts
from experimentally reared C. decemmaculatus.
The elongate body measures 1,728–2,256 (2,026)×208–
256 (239) at the acetabular level. The tegument is spinous,
with scale-like spines 10–13 long arranged in dense rows in
the forebody and extended up to the ventral sucker dorsally
and in lateral bands ventrally (Fig. 3i–j); in the hindbody,
the spines cover the ventral surface, decrease in size and
density and reach a short distance behind the testes. The
dorsal surface is free from spines, except for the narrow
lateral bands.
The small head collar, 158–189 (172) wide, is armed
with 22 straight spines (Fig. 2b) arranged in a single,
dorsally interrupted row and distributed as follows: two
small corner spines, the first one measuring 23.7–31.6
(26.0; 2.1 n=20) long and the second one 26.9–33.2 (29.3;
1.6 n=19) long, the next spine on each side is slightly
Parasitol Res (2008) 102:647–655
encysted on gill of Moenckhausia dichroura, scale 100 μm; forebody
of adult: i dorsal side; j ventral side, scale 100 μm; k adult, anterior
level of vitelline follicles (arrows), scale 200 μm
larger than the corner spines, 26.9–33.2 (30.4; 1.9; n=19)
long, but smaller than the subsequent four spines, 28.4–
34.8 (31.8; 1.4; n=36) long; last four dorsal spines 28.4–
37.9 (33.8; 3.1; n=11) long.
The terminal oral sucker, 60–88 (76)×63–82 (73), is
armed with ten small spines on the anterior border
(Fig. 2b); the prepharynx is short, 0–47 (22); the pharynx
is muscular, 69–95 (80)×63–85 (67); the long oesophagus
161–221 (189) bifurcates a short distance anterior to the
ventral sucker into two caeca, which end in the distal
chamber of the excretory vesicle forming the uroproct. The
pharynx to oral sucker width ratio is 1:1.0–12 (1.1). The
ventral sucker, 142–173 (154)×158–180 (171), is placed in
the anterior third of the body and bears about 30 small
spines or scales around the opening. The oral sucker to
ventral sucker width ratio is 1:2.1–2.7 (2.3).
The genital organs are found in the middle third of the
body. The oval testes are generally in contact and arranged
in tandem; the anterior testis, 117–189 (158)×139–189
(160), is slightly shorter and wider than the posterior
Parasitol Res (2008) 102:647–655
testis, 123–261 (222)×139–176 (145), and both testes
show regular borders. The cirrus sac, 120–176 (142)×95–
113 (103), is dorsal to and partly covered by the ventral
sucker. The seminal vesicle is bipartite, the prostatic
glands are visible, and the inconspicuous cirrus is short.
The genital pore is situated anterior to the ventral sucker.
The round ovary, 54–95 (72)×79–117 (97), is located in
front of the anterior testis; the Mehlis’ gland and the
uterine seminal receptacle are present. The short uterus,
which contains up to 15 eggs 88–104 (94)×35–60 (49),
occupies the space between the anterior testis, and the
ventral sucker and ends in a delicate duct at the genital
pore. The distance between the posterior border of the
ventral sucker and the anterior border of the ovary
represents 6.7–9.4% (8.0%) of the total body length
(TBL). The vitelline glands, which are transversally
elongate and relatively large, 19–38 (28)×25–101 (53),
are generally extended in two lateral bands from various
points of the anterior testis to the body end, sometimes
reaching even up to the level of the anterior border of
ovary (Fig. 3k); they become more or less confluent
behind the testes. The excretory vesicle possesses five
chambers: a shorter distal chamber receiving the intestinal
caeca and four larger anterior chambers, with collecting
ducts arising from the proximal chamber at each body
side. The post-testicular space represents 31.8–43.1%
(38.1%) of the TBL.
Five of eight chicks previously force-fed with cysts
became positive. Three of them, infected each with 20, 40
and 50 cysts from experimentally reared C. decemmacula-
tus, yielded 1, 22 and 39 parasites at 3, 5 and 7 days pe,
respectively. Two chicks, each of which was force-fed with
30 cysts from exposed C. decemmaculatus of natural
source, resulted infected with five and six parasites at
day 7 pe. The remaining three chicks infected (1) with eight
cysts from P. reticulata and (2) with 11 and 36 cysts from
exposed C. decemmaculatus of natural source were
negative.
Eggs and miracidia
Eggs in freshly passed faeces of chicks are 91–104 (95)×
54–60 (57; n=30). Each egg is surrounded by an oval
capsule, yellow to light brown, operculate and contains a
zygote and several vitelline cells. At 26–28°C, the eyespots
of miracidia appeared on day 10; miracidia became fully
developed between 13 and 15 days and emerged 3 days
later (Fig. 3a–c). Live miracidia are up to 125 long, but
fixed specimens measure 60–74 (68)×35–46 (39; n=16).
Miracidia swam actively during 24 h, and swimming
slowed down for the next 48 h. The maximal life span
was about 4 days, but infection probably took place within
the first 24 h at room temperature (13–25°C).
651
Sporocysts
The sporocysts are found on the gill of the snail; live
sporocysts measure 230–340×220–340 (n=3; 28 days pe)
and 527×232 (n=1; 43 days pe). The body is brown
pigmented and remnants of miracidial eyespot pigment are
still visible; one to two mother rediae 148–251×60–102,
with a pharynx 34–55×29–49 can be seen inside the
sporocyst. No sporocysts were detected in the snails
dissected at 71 and 136 days pe (Fig. 3d–e).
Rediae
Measurements were made on permanent mounted speci-
mens (Figs. 2c–e and 3f).
The first two free mother rediae 553–565×188, with a
pharynx 72–79×79 were found on the gill 43 days pe
(Fig. 2c). They show a feeble tegumental collar behind the
pharynx, a pair of short appendages near the posterior end
of the body; the caecum reaches to the level of the
appendages. The body cavity contains about seven large
and some smaller germ balls.
At day 72 pe, there were about 50 rediae invading the
gill, hepatopancreas and gonad. The smallest rediae
measured 142–283×25–50 with a pharynx 16–38×19–35
(n=23), whereas the largest ones were 314–710×44–157
with a pharynx 25–126×38–126 (n=16); 12 of the latter
contained small daughter rediae (Fig. 3f).
At day 136 pe, the number of rediae increased up to 250;
rediae were variable in size, measuring 226–735×50–144,
with a pharynx 19–95×19–100 (n=27); 12 of the measured
rediae contained small daughter rediae (Fig. 2e).
All rediae containing daughter rediae are larger than 330;
the small daughter rediae are 100–190×28–47, with a small
pharynx 16–28×19–32, similar in size to the smallest free
rediae. No developing cercariae can be distinguished at this
time. At least three pairs of protonephridia are present in
living mother rediae.
At day 240 pe, an uncounted number of daughter rediae
with developing cercariae had invaded the entire snail.
Daughter rediae are morphologically similar to mother
redia, but larger, 389–810 (612)×82–220 (136), show a
smaller pharynx 44–66 (50) x 47–57 (52; n=12), a less
conspicuous annular collar in fixed specimens, two poste-
rior appendages, and the caecum reaches to the level of
posterior appendages. They differ from mother rediae in
that they contain immature cercariae, which are clearly
distinguished by the presence of oral and ventral suckers, a
short tail and undeveloped germ balls. Immature cercariae
leave the rediae and complete their development in the
tissues of the snail.
The pharynx size (69–126×82–126) in 7 of the 24 rediae
containing small daughter rediae was comparable to that of
652
two rediae found free at day 43 pe; the pharynx size of the
remaining rediae (35–66×44–69) was similar to that of
daughter rediae (44–66×47–57) with developing cercariae
at day 240 pe. This fact suggests the occurrence of more
than two generations of rediae or that daughter rediae are
capable to produce rediae before they switch to cercarial
production.
The three snails fixed on 29, 58, and 111 days pe for
histological sections show the sporocyst or the rediae
invading the snails in a similar manner as the snails
dissected 42, 72 and 136 days pe, respectively.
Rediae from natural infections (based on 15 formalin-
fixed unstained specimens mounted under coverslip without
pressing on it) varied considerably in body size, 560–1136
(838)×160–240 (201) and showed a muscular pharynx 38–
94 (71)×38–88 (66; Fig. 2d).
All the eight exposed A. chloroticum specimens became
infected, whereas none of the H. parchappei snails was
positive after dissection between weeks 4 and 7 pe.
Cercariae
Measurements were based on 20 fixed cercariae from
naturally infected snails (Figs. 2f–g and 3g). The body
239–264 (250)×82–94 (88) has a tegument without spines.
The head collar is feebly developed and lacks spines. The
oral sucker is subterminal, 35–47 (39)×38–44 (40), and has
ten flattened spines on its anterior border. The prepharynx
is 25–32 (28); there is a small prepharyngeal body, 9–16
(14)×9–13 (12), with two globular inclusions. The pharynx
is muscular, 16–25 (20)×13–16 (14), the oesophagus is
long, and the distance between the oral and ventral suckers
is 126–145 (135); the intestine bifurcates shortly anterior to
the ventral sucker, and the caeca reach to the level of the
excretory vesicle. The ventral sucker 41–47 (46)×38–44
(42), which is slightly larger than the oral one, is situated
posterior to the midbody and shows flattened spines
surrounding the opening. Cystogenous cells with bar-
shaped contents lie between the pharynx and the end of
the body. Penetration glands, gland ducts openings anterior
to oral sucker and paraoesophageal gland cells are not
observed. The genital rudiments consist of two masses of
cells, one at the anterior and the other at the posterior
margin of the ventral sucker; the cell masses are connected
by a string of cells passing dorsal to the acetabulum.
The excretory system is stenostomate, with the main
tubes extending from the anterior wall of the small
excretory vesicle to the level of the prepharynx; the main
collecting tubes are narrow, with 23–51 refractile granules
measuring 5–6 in diameter. The excretory ducts run
posteriad after forming a loop at the prepharyngeal level;
the bifurcation into the anterior and posterior canals is not
clearly seen, but would occur at the posterior level of the
Parasitol Res (2008) 102:647–655
ventral sucker, as indicated by ciliary patches distributed
along the interior margins. The flame cells are arranged in
16 pairs (Fig. 2f). In emerged cercariae, the caudal duct of
the excretory system enters the anterior portion of the tail
where it ends blindly after a short distance. In immature
cercariae within snails, the caudal duct of the excretory
system runs along the tail, bifurcates into two short
branches near the posterior end and opens at a pore. As
the cercaria matures, the short branches become blinded,
and the whole duct shortened.
The tail is 1,792–2,224 (1,919)×138–192 (161) and not
pigmented. Longitudinal muscle fibres run along the central
axis of the tail; circular muscles, arranged in packets, give a
characteristic feature (Fig. 2f–g)
At room temperature (13–25°C), cercariae survived up
to 4 days. They swam actively while describing an S-like
figure for approximately 24 h and then floated for a short
time in stretched position as they fell to the bottom where
the tail coiled over the body (Fig. 3g). The cercariae lost
vitality during the following 2 days keeping the tail in this
position. In the last period of life until death, they were
unable to coil the tail. At 26–28°C cercarial longevity was
shorter, with 50% of them active during 24 h and a
maximum survival of 48 h.
No penetration of fish by cercariae was observed, but
they may have been swallowed or may have reached the
gill filaments passively with the respiratory current.
Experimentally obtained cercariae were about 10%
smaller in body and tail size than naturally obtained
cercariae. The last surviving snail emitted the first cercariae
at 6 months pe and continued emitting for about more than
2 months. The snail was dissected after cercarial shedding
ceased.
Encysted metacercaria
Measurements were based on ten live metacercariae (obtained
between 10 and 15 d pe) encysted on the gill filaments of
experimentally infected C. decemmaculatus; the measure-
ments of ten specimens from M. dichroura are in brackets
(Fig. 3h). The oval cysts, 120–136 (127)×95–113 (102)
[117–126 (120) x 95–104 (99)], have a thin wall. The
corpuscles in the excretory system are conspicuous, 3–6 (5)
in diameter [5–11 (7)×3–8 (6)]. The head collar bears a
dorsally interrupted crown of 22 oral spines, with corner
spines 9.5–11.7 (11); the first spine of the row is 9.5–10.6
(10) and the next spines are 10.6–13.8 (12) long. The head
collar spines became visible from day 10 pe onward. There
are six to eight pores at the front of the oral sucker, which
presumably are penetration gland outlets. The oral sucker is
30–36 (34)×36–44 (42) [25–35 (30)×40–47 (42)] and the
ventral sucker 32–41 (35)×47–52 (51) [28–43 (35)×47–63
(54)]. The prepharynx is short, the pharynx is oval, 19–27
Parasitol Res (2008) 102:647–655
(23)×13–16 (14) [17–25 (22)×13–24 (18)], and the intesti-
nal caeca are not visible due to the small size of the cysts.
Although P. reticulata was easily infected with cercariae,
only a few cysts of metacercariae were present between 2
and 3 weeks pe. The cysts survived for at least 2 months in
C. decemmaculatus.
The fishes caught at the same site in Yacyretá dam where
the infected snails were collected showed a high prevalence
but low intensity (2–24) of infection with cysts of S.
aylacostoma in the gills: M. dichroura 88.9% (n=9), A.
erythropterus 100% (n=8), H. serpae 100% (n=6) and
undetermined juveniles 85.7% (n=7). A group of 14 fishes
belonging to the same species caught from a site without
Aylacostoma snails were free of cysts.
Taxonomic summary
Natural definitive unknown
host
Experimental Gallus gallus f. domestica (L.)
definitive host
Site of infection posterior half of intestine
Etymology the species is named after the genus of
its first intermediate host.
Specimens MACN-Pa 441/1-6
deposited
First intermediate Aylacostoma chloroticum Hylton Scott,
host 1954 (Thiaridae)
Second intermediate Cnesterodon decemmaculatus (Jenyns
host 1842), Poecilia reticulata (Peters
1859) (Poeciliidae; experimental);
Moenckhausia dichroura (Kner 1858),
Astyanax erythropterus (Holmberg
1899), Hyphesobrycon serpae (Durbin
in Eigenmann 1908)
(Tetragonopteridae; natural).
Metacercariae encysted on gill
filaments and gill chamber.
Locality Paraná River in Heller Peninsula, near
Posadas City, Province of Misiones,
Argentina.
Discussion
This is the first report of a species of Aylacostoma infected
with larval stages belonging to a new trematode species
whose life cycle was experimentally established.
Stephanoprora aylacostoma n.sp. differs from other
species of the genus in its larger eggs, in the smaller,
slender body and smaller collar spines of the adult and in
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the morphological and biological features of the larval
stages.
The life cycle of the new species is similar to that of S.
denticulata from Europe in that prosobranch snails and
fishes act as intermediate hosts and in having large-tailed
cercariae and metacercariae which encyst on fish gills (Køie
1986). The cercaria of S. aylacostoma differs from that of S.
denticulata in possessing a prepharyngeal body and
corpuscles in the excretory duct, in lacking a long excretory
duct in the tail and in the behaviour during the active
swimming and resting phases. Although Køie (1986) did
not describe the mature adults obtained experimentally
between 7 and 10 days pe, she kindly sent us a specimen
for comparison. This adult differs from S. aylacostoma in
its larger body size (3,568×320), smaller eggs (90×42) and
the extension of the tegument spines.
S. aylacostoma is morphologically similar to S. dentic-
ulata (Rud.) described by Nasir and Scorza (1968) and to S.
paradenticulata described by Nasir et Rodriguez (1969),
both from Venezuela, and differs from them in its smaller
body and collar spines and in possessing larger eggs. The
life cycle of S. denticulata includes a pulmonate snail and a
cercaria with collar spines and 18 pairs of flame cells,
which encysts on the inner surface of the oesophagus of the
experimental fishes (Nasir and Scorza 1968). The cercaria
of S. paradenticulata possesses 18 flame cells and a tail as
long as the body (Nasir et Rodriguez 1969). However, the
life cycle and taxonomic status of S. denticulata as reported
by Nasir and Scorza (1968) may be subject to revision
because differences with the European S. denticulata
suggest that it has been misclassified (Køie 1986) or that
experimental results were misinterpreted (Ostrowski de
Núñez 2007).
S. aylacostoma is distinguished from the experimentally
obtained adults of S. uruguayense of the same age
(Ostrowski de Núñez 2007) only by its smaller body, tail
and collar spines and larger eggs. The main differences
between these species lay in the morphological and
biological features of life cycle stages and in the interme-
diate hosts. S. aylacostoma exhibits the following distinc-
tive features in comparison with S. uruguayense: adult body
size is smaller (2,026 vs 2,770), live eggs are larger (95×57
vs 83×51), miracidia are larger (68 vs 52) and have a
considerably longer life span (4 days vs 20 h); daughter
rediae from natural infections are shorter and wider (836×
201 vs 976×150), and the pharynx is larger (71×66 vs 44×
50); the cercariae are larger (250 vs 183) and possess
calcareous corpuscles in the excretory system, which are
absent in the cercariae of S. uruguayense; the metacercaria
cysts are larger (127×120 vs 108×69), the calcareous
corpuscles are smaller (7×6 vs 17×9), and the head collar
spines appear later (10 days vs 7 days) (Ostrowski de
Núñez 2007). The first intermediate host of S. aylacostoma
654
seems to be specific, as its miracidia are unable to infect H.
parchappei, the host of S. uruguayense. The morphological
features of S. aylacostoma fit well with that stated as typical
for species of Stephanoprora by Ostrowski de Núñez
(2007) except for the presence of corpuscles in the
excretory system of the large-tailed cercaria.
The other macrocercous cercariae from prosobranch snails
of unknown life cycle, which are comparable to the present
form are Cercaria illecebrosa Lee and Seo 1959 from
Amnicola limosa (Say) in Michigan (USA) and Cercaria
heleobicola IV Martorelli, 1990 from Heleobia conexa
(Gaillard 1974) in Argentina. Both cercariae differ from that
of S. aylacostoma mainly in lacking corpuscles in the
excretory system and in the absence of a prepharyngeal body.
One should be cautious when comparing S. aylacostoma
with other described species from birds in South America,
taking into account that data were only obtained from
adults recovered at day 7 pe. Furthermore, Stephanoprora
species are distinguished by just a few characters and may
survive in the host for at least 30 days while the body,
organs and collar spines increase in size (Ostrowski de
Núñez et al. 2004). Although Stephanoprora species may
be difficult to distinguish from each other at the adult stage,
they differ in the morphological features of the larval stages
and biological characteristics of the life cycle.
Stephanoprora aylacostoma is similar in size to S.
podicipei Etchegoin et Martorelli, 1997, which parasitised
Podiceps major and Sterna hirudinacea from a marine
habitat located near the Atlantic Ocean, in the south of
Buenos Aires Province (Etchegoin and Martorelli 1997;
Cremonte et al. 1999).
Stephanoprora podicipei was ascribed to S. uruguayense
by Ostrowski de Núñez et al. (2004) based on the
similarities in morphology and measurements. The further
knowledge of the life cycle of S. podicipei—with a
probable marine first intermediate host—will allow to
determine whether it is a separate species to S. uru-
guayense; the difference to S. aylacostoma is given by the
specific freshwater intermediate host with its restricted
distribution in the Paraná River.
On the other hand, snails of the genus Aylacostoma are
distributed over a wide region of tropical South America
(Venezuela, Colombia, Peru, Brazil) and may be involved
in the life cycle of S. conciliata Dietz 1910. This species
was described based upon specimens collected by J.
Natterer near Rio de Janeiro, Brazil (1817–1835) and later
mentioned by Lutz (1928) who provided vague and
inaccurate information on the life cycle. S. conciliata is
similar to S. aylacostoma in the size of the body (1.4–
2.1 mm) and collar spines (corner spines 21 long,
subsequent spines 29–38), but possesses smaller eggs
(65–67 × 43–45) and generally smaller organs (Dietz
1910). At present, the two species are considered different,
Parasitol Res (2008) 102:647–655
but classification may change with further knowledge of the
life cycle of S. conciliata and with an adequate redescrip-
tion based upon fresh material from the type host.
Stephanoprora argentinensis (Sutton et al. 1982) inhab-
iting Patagonian lakes, differs clearly from the new species
in constantly having 20 collar spines, larger body (up to
3,318) and smaller eggs (64–76×36–56).
S. aylacostoma is considered a new species based on the
distinct morphology of the adult and on the morphological
and biological characteristics of the life cycle stages
mentioned above. In the near future, the filling up of the
Yacyretá reservoir will modify dramatically the ecology of
the river, threatening the existence of A. chloroticum in the
subtropical region of Argentina and probably preventing
this trematode from completing the life cycle.
Acknowledgements We are indebted to the late Dr. C. Sutton
(Universidad Nacional de La Plata) for the loan of paratypes; to Dr. P.
Pillit (United States Department of Agriculture) for the loan of
vouchers and type material; to Dr. F.Cremonte (Centro Nacional
Patagónico, Pto. Madryn, Argentina) and Dr.M.Køie (Marine Biolog-
ical Laboratory, Helsingør, Denmark) for the loan of specimens from
their personal collections; to Mr. P.E. Topa (CEPAVE, La Plata,
Argentina) for his valuable assistance in the preparation and
processing of histological slides of the molluscs and to Lic. D.C.
Pérez (Entidad Binacional Yacyretá) and Lic. J.G. Peso (Universidad
Nacional de Misiones) for their help in the collection of snails.
Funds were provided by CONICET Grant (BID 1201/OC-AR-PICT
no.1-6604) and by EBY (Yacyretá Binational Entity) to the Aylacos-
toma Program in Yacyretá.
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